摘要
目的探讨血管紧张素Ⅱ受体1型(AT1R)—半胱氨酸丰富跨膜成骨蛋白调控因子(Crim1)信号通路在乳鼠肥大心室肌细胞编码内向整流钾电流离子通道的Kir2.1 mRNA和蛋白表达调控中的作用。方法分离1 d龄SD大鼠乳鼠心室获心室肌细胞,给予腺病毒空载体(Ad-Null)、Crim1基因重组腺病毒载体(Ad-Crim1)、血管紧张素Ⅱ(AngⅡ)和/或氯沙坦(Los)分组干预。实时荧光定量逆转录PCR检测肌球蛋白重链β(β-MHC)、Kir2.1和Crim1的mRNA表达。Western免疫印迹法检测全细胞提取蛋白Kir2.1和Crim1的表达,细胞结晶紫染色测量表面积。结果AngⅡ干预24 h明显增加心室肌细胞β-MHC的mRNA表达及细胞面积;下调Crim1和Kir2.1的mRNA和蛋白表达。Ad-Crim1和Los干预明显抑制AngⅡ干预的上述效应;Ad-Crim1联合Los较Ad-Crim1单独应用能更显著抑制AngⅡ刺激的上述效应。结论AT1R-Crim1信号通路参与乳鼠肥大心室肌细胞Kir2.1 mRNA和蛋白表达的调控。
Objective To investigate the regulation of angiotensinⅡtype 1 receptor(AT1R)-Cysteine-rich transmembrane bone morphogenetic protein regulator 1(Crim1)signaling pathway on the expression of Kir2.1 of inward rectifier potassium current ion channel at mRNA and protein levels in the hypertrophic ventricular myocytes from neonatal rats.Methods The ventricular myocytes were isolated from the ventricles of 1 d-old neonatal Sprague-Dawley rats and intervened with adenovirus empty vector(Ad-Null),Crim1 gene recombinant adenovirus vector(Ad-Crim1),AngiotensinⅡ(AngⅡ)and/or losartan(Los).The mRNA expression of myosin heavy chain beta(β-MHC),Crim1and Kir2.1were detected by RT-qPCR.The protein levels of Crim1 and Kir2.1 in the whole-cell extracts were determined by western blot(WB)analysis.Cell surface area was measured by crystal violet staining.Results Treatment of the neonatal rat ventricular myocytes with AngⅡfor 24 h increased the mRNA expression ofβ-MHC and the size of the cell surface.Treatment of the cells with AngⅡdecreased the mRNA and protein expressions of Crim1 and Kir2.1.Both Ad-Crim1 and Los prevented those effects of AngⅡintervention.Compared with Ad-Crim1 alone,Ad-Crim1 combined with Los can significantly inhibit the above effects of AngⅡstimulation.Conclusion AT_(1)R-Crim1 signaling pathway participates in regulating the mRNA and protein expressions of Kir2.1 in the hypertrophic ventricular myocytes from neonatal rats.
作者
霍照美
杨英
杨龙
何炯红
夏桂玲
郭楚娴
Huo Zhaomei;Yang Ying;Yang Long;He Jionghong;Xia Guiling;Guo Chuxian(Guizhou Medical University,Guiyang 550025,Guizhou,China;Department of Cardiology,Guizhou Provincial People’s Hospital,Guiyang 550002,Guizhou,China)
出处
《贵州医药》
CAS
2022年第3期339-342,共4页
Guizhou Medical Journal
基金
贵州省科技计划项目[黔科合基础(2019)1205]
贵州省科学技术基金[黔科合基础-ZK(2021)一般360]
贵州省科技平台及人才团队计划[黔科合平台人才(2017)5405]。
关键词
心肌肥大
室性心律失常
钾离子通道
血管紧张素Ⅱ受体1型
半胱氨酸丰富跨膜成骨蛋白调控因子
Cardiac hypertrophy
Ventricular arrhymaias
Potassium ion channel
AngiotensinⅡtype 1 receptor
Cysteine-rich transmembrane bone morphogenetic protein regulator 1
